In the following discussion certain articles and methods will be described for background and introductory purposes. Nothing contained herein is to be construed as an admission of prior art. Applicant expressly reserves the right to demonstrate, where appropriate, that the articles and methods referenced herein do not constitute prior art under the applicable statutory provisions.
Fibrinogen is a key player in the coagulation cascade and has critical functions in the regulation of inflammation (Davalos and Akassoglou, Semin. Immunopathol. 2012 34(1):43-62). Although soluble fibrinogen in the bloodstream is not proinflammatory, its conversion to insoluble fibrin after activation of coagulation leads to exposure within the fibrinogen y chain of the cryptic epitope γ377_395, which binds to CD11b/CD18 integrin (Mac-1, complement receptor 3, aM˜2) (Altieri et al., J Biol Chem. 1990 Jul. 25; 265(21):12119-22; Lishko et al., 2002; Biochemistry. 2002 Oct. 29; 41(43):12942-51.). Epitope exposure can occur during fibrin formation in vivo, or upon fibrinogen immobilization on substrates. However, there is no method described to efficiently reproduce the in vivo configurations of fibrin provided by its interactions with other molecules in vitro, or to predict the functional activity of an agent that binds to fibrin based on an in vitro assay.
There is thus a need in the art for a method of preparing fibrin compositions with the ability to predict functional utility using activity demonstrated in an in vitro setting. The present invention addresses this need.